Inhibition of apoptosis process and improvement of cell performance
View Patent ↗The present invention relates to preventing or delaying programmed cell death by expressing one or more anti-apoptotic polypeptides in a cell such that programmed cell death in the cell is prevented or delayed. The present invention also relates to increasing production of a cell-related product by expressing one or more anti-apoptotic polypeptides in a cell such that production of the cell-related product by the cell is increased. Recombinant cells useful for producing cell-related product or cellular therapy are also provided.
1. A method for preventing or delaying programmed cell death in an isolated recombinant cell, the method comprising expressing two or more anti-apoptotic polypeptides in the cell such that programmed cell death in the cell is prevented or delayed, wherein the two or more anti-apoptotic polypeptides comprise E1B-19K and Aven, and wherein the cell further comprises heterologous polynucleotides encoding E1B-19K and Aven.
2. The method of claim 1 , wherein expression of two or more anti-apoptotic polypeptides is controlled by at least one inducible heterologous promoter operably linked to a the heterologous polynucleotides encoding E1B-19K and Aven.
3. The method of claim 2 , wherein the inducible heterologous promoter is inducible by a steroid.
4. The method of claim 2 , wherein the inducible heterologous promoter is an ecdysone promoter.
5. The method of claim 2 , wherein the isolated recombinant cell further comprises a screenable or selectable marker operably linked to the at least one inducible heterologous promoter.
6. The method of claim 5 , wherein the screenable or selectable marker is green fluorescence protein (GFP) or enhanced green fluorescence protein (EGF).
7. The method of claim 1 , wherein the anti-apoptotic polypeptides are encoded by genes in eukaryotic cells or viruses.
8. The method of claim 1 , wherein the isolated recombinant cell is a mammalian cell.
9. The method of claim 8 , wherein the isolated recombinant cell is selected from the group consisting of human cells, murine cells, and rodent cells.
10. The method of claim 8 , wherein the isolated recombinant cell is a CHO cell.
11. The method of claim 8 , wherein the isolated recombinant cell is a BHK cell.
12. The method of claim 1 , wherein the isolated recombinant cell is in a large scale bioreactor or culture device of a commercial production.
13. A method of increasing production of a cell-related product by an isolated recombinant cell, the method comprising expressing two or more anti-apoptotic polypeptides in the cell such that production of the cell-related product by the cell is increased, wherein the two or more anti-apoptotic polypeptides comprise E1B-19K and Aven, and wherein the cell further comprises heterologous polynucleotides encoding E1B-19K and Aven.
14. The method of claim 13 , wherein the cell-related product is selected from the group consisting of cell-secretion factors.
15. The method of claim 13 , wherein expression of two or more anti-apoptotic polypeptides is controlled by at least one inducible heterologous promoter operably linked to the heterologous polynucleotides encoding E1B-19K and Aven.
16. The method of claim 15 , wherein the inducible heterologous promoter is inducible by a steroid.
17. The method of claim 15 , wherein the inducible heterologous promoter is an ecdysone promoter.
18. The method of claim 15 , wherein the isolated recombinant cell further comprises a screenable or selectable marker operably linked to the at least one inducible heterologous promoter.
19. The method of claim 18 , wherein the screenable or selectable marker is green fluorescence protein (GFP) or enhanced green fluorescence protein (EGF).
20. The method of claim 13 , wherein the anti-apoptotic polypeptides are encoded by genes in eukaryotic cells or viruses.
21. The method of claim 13 , wherein the isolated recombinant cell is a mammalian cell.
22. The method of claim 21 , wherein the isolated recombinant cell is selected from the group consisting of human cells, murine cells, and rodent cells.
23. The method of claim 21 , wherein the isolated recombinant cell is a CHO cell.
24. The method of claim 21 , wherein the isolated recombinant cell is a BHK cell.
25. The method of claim 13 , wherein the isolated recombinant cell is in a large scale bioreactor or culture device of a commercial production.
26. A method of increasing production of an isolated recombinant cell product, the method comprising expressing two or more anti-apoptotic polypeptides in the cell such that production of the isolated recombinant cell product is increased, wherein the two or more anti-apoptotic polypeptides comprise E1B-19K and Aven, and wherein the cell further comprises heterologous polynucleotides encoding E1B-19K and Aven.
27. An isolated recombinant cell useful for producing cell-related product expressing two or more anti-apoptotic polypeptides, wherein the two or more anti-apoptotic polypeptides comprise E1B-19K and Aven, and wherein the cell further comprises heterologous polynucleotides encoding E1B-19K and Aven.
28. The isolated recombinant cell of claim 27 , wherein the anti-apoptotic polypeptides are encoded by genes in eukaryotic cells or viruses.
29. The isolated recombinant cell of claim 27 , wherein the cell-related product is selected from the group consisting of cell-secretion factors.
30. The isolated recombinant cell of claim 27 , wherein expression of two or more anti-apoptotic polypeptides is controlled by at least one inducible heterologous promoter operably linked to the heterologous polynucleotides encoding E1B-19K and Aven.
31. The isolated recombinant cell of claim 30 , wherein the inducible heterologous promoter is inducible by a steroid.
32. The isolated recombinant cell of claim 30 , wherein the inducible heterologous promoter is an ecdysone promoter.
33. The isolated recombinant cell of claim 30 , wherein the isolated recombinant cell further comprises a screenable or selectable marker operably linked to the at least one inducible heterologous promoter.
34. The isolated recombinant cell of claim 33 , wherein the screenable or selectable marker is green fluorescence protein (GFP) or enhanced green fluorescence protein (EGF).
35. The isolated recombinant cell of claim 27 , wherein the isolated recombinant cell is a mammalian cell.
36. The isolated recombinant cell of claim 27 , wherein the isolated recombinant cell is selected from the group consisting of human cells, murine cells, and rodent cells.
37. The isolated recombinant cell of claim 35 , wherein the isolated recombinant cell is a CHO cell.
38. The isolated recombinant cell of claim 27 , wherein the isolated recombinant cell is a BHK cell.
39. The isolated recombinant cell of claim 27 , wherein the cell produces cell-related product in a large scale bioreactor or culture device of commercial production.
40. A population of isolated recombinant cells useful for producing one or more biological functions of the cells expressing two or more anti-apoptotic polypeptides, wherein the two or more anti-apoptotic polypeptides comprise E1B-19K and Avena and wherein the cell further comprises heterologous polynucleotides encoding E1B-19K and Aven.
41. An isolated recombinant cell with prevented or delayed programmed cell death, prepared by the method of claim 1 .
42. The method of claim 1 , wherein the isolated recombinant cell is derived from a single cell.
43. The method of claim 13 , wherein the isolated recombinant cell is derived from a single cell.
44. The method of claim 26 , wherein the isolated recombinant cell is derived from a single cell.
45. The isolated recombinant cell of claim 27 , wherein the isolated recombinant cell is derived from a single cell.
46. The isolated recombinant cell population of claim 40 , wherein the isolated recombinant cell population is derived from a single cell.